Fast sequential multi-element determination of Ca, Mg, K, Cu, Fe, Mn and Zn for foliar diagnosis using high-resolution continuum source flame atomic absorption spectrometry: Feasibility of secondary lines, side pixel registration and least-squares background correction

The fast sequential multi-element determination of Ca, Mg, K, Cu, Fe, Mn and Zn in plant tissues by high-resolution continuum source flame atomic absorption spectrometry is proposed. For this, the main lines for Cu (324.754 nm), Fe (248.327 nm), Mn (279.482 nm) and Zn (213.857 nm) were selected, and the secondary lines for Ca (239.856 nm), Mg (202.582 nm) and K (404.414 nm) were evaluated. The side pixel registration approach was studied to reduce sensitivity and extend the linear working range for Mg by measuring at wings (202.576 nm; 202.577 nm; 202.578 nm; 202.580 nm: 202.585 nm; 202.586 nm: 202.587 nm; 202.588 nm) of the secondary line. The interference caused by NO bands on Zn at 213.857 nm was removed using the least-squares background correction. Using the main lines for Cu, Fe, Mn and Zn, secondary lines for Ca and K, and line wing at 202.588 nm for Mg, and 5 mL min(-1) sample flow-rate, calibration curves in the 0.1-0.5 mg L-1 Cu, 0.5-4.0 mg L-1 Fe, 0.5-4.0 mg L-1 Mn, 0.2-1.0 mg L-1 Zn, 10.0-100.0 mg L-1 Ca, 5.0-40.0 mg L-1 Mg and 50.0-250.0 mg L-1 K ranges were consistently obtained. Accuracy and precision were evaluated after analysis of five plant standard reference materials. Results were in agreement at a 95% confidence level (paired t-test) with certified values. The proposed method was applied to digests of sugar-cane leaves and results were close to those obtained by line-source flame atomic absorption spectrometry. Recoveries of Ca, Mg, K, Cu, Fe, Mn and Zn in the 89-103%, 84-107%, 87-103%, 85-105%, 92-106%, 91-114%, 96-114% intervals, respectively, were obtained. The limits of detection were 0.6 mg L-1 Ca, 0.4 mg L-1 Mg, 0.4 mg L-1 K, 7.7 mu g L-1 Cu, 7.7 mu g L-1 Fe, 1.5 mu g L-1 Mn and 5.9 mu g L-1 Zn. (C) 2009 Elsevier B.V. All rights reserved.

Rabies laboratory diagnosis: peculiar features of samples from equine origin

O diagnóstico laboratorial da raiva é realizado através de métodos de pesquisa do corpúsculo de Negri, imunofluorescência direta e inoculação em camundongos. Na maioria dos casos, quando a amostra é bem coletada, bem conservada e o profissional responsável possui experiência, verifica-se concordância entre as técnicas utilizadas. A Seção de Raiva e Encefalomielite do Instituto Biológico ao comparar a sensibilidade das três técnicas diagnósticas, em 3713 amostras (córtex cerebral, cerebelo e hipocampo) recebidas no período de 1980-1994, sendo 3010 da espécie bovina (983 positivas) e 703 da espécie eqüina (111 positivas), observou que, no caso da raiva eqüina, esta concordância não é mantida. Verificou-se, nesta espécie, que somente em algumas oportunidades foi possível identificar, pelo método histopatológico, o corpúsculo de Negri. em relação à prova de imunofluorescência pode-se afirmar que a mesma detectou uma porcentagem menor de amostras positivas, provenientes da espécie equina, em compração com as da espécie bovina, sendo esta diferença estatisticamente significativa. A prova biológica foi a mais sensível, porém houve uma diferença, também significativa, entre o período de incubação em camundongos das amostras de origem bovina e das de origem eqüina. A presença pouco frequente de corpúsculos de Negri e o período de incubação em camundongos mais prolongado, das amostras de origem eqüina, sugerem que devem ser intensificados os estudos da patogenia da raiva nesta espécie.

Comparison of histological and molecular diagnosis of Helicobacter pylori in benign lesions and gastric adenocarcinoma

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Interference of vaccinal antibodies on serological diagnosis of leptospirosis in vaccinated buffalo using two types of commercial vaccines

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytotaxonomic diagnosis of Trichomycterus diabolus (Teleostei: Trichomycteridae) with comments about its evolutionary relationships with co-generic species

São descritos o cariótipo e a localização das regiões organizadoras de nucléolo (Ag-NOR) de uma amostra de Trichomycterus diabolus, coletada no córrego Hortelã (Botucatu, São Paulo, Brasil). A espécie apresentou 2n=56 cromossomos (42 metacêntricos, 12 submetacêntricos e 2 subtelocêntricos) e as regiões organizadoras de nucléolo localizadas próximas ao centrômero, no braço longo do maior par metacêntrico. A ocorrência de 2n=56 cromossomos em Trichomycterus diabolus é uma característica interessante, uma vez que, até o momento, todas as espécies cis-Andinas cariotipadas apresentaram 2n=54 cromossomos, enquanto que quase todas as espécies trans-Andinas apresentaram números diplóides diferentes. É discutida a possível origem desta inesperada estrutura cariotípica.

Oral lichen planus versus epithelial dysplasia: difficulties in diagnosis

O diagnóstico histopatológico do líquen plano bucal não é fácil, pois alguns casos de displasia epitelial podem apresentar características bastante semelhantes às do líquen plano. OBJETIVO: Comparar as alterações celulares sugestivas de malignidade presentes no líquen plano bucal com as da displasia epitelial. MATERIAL E MÉTODOS: Cortes histológicos de líquen plano bucal e displasia, corados com hematoxilina-eosina, foram analisados por meio da microscopia de luz. RESULTADOS: A análise de variância (alfa=5%) revelou haver diferença estatisticamente significante entre o número médio de alterações celulares no líquen plano bucal (5,83±1,61) e na displasia epitelial (4,46±1,26). O teste de qui-quadrado não mostrou diferença estatisticamente significante entre o líquen plano bucal e a displasia epitelial em relação às seguintes alterações celulares: aumento da relação núcleo/citoplasma, hipercromatismo nuclear, distribuição irregular da cromatina e núcleos aumentados (p>0,05). CONCLUSÃO: Algumas alterações celulares sugestivas de malignidade presentes no líquen plano bucal também podem ser encontradas na displasia epitelial, dificultando o seu diagnóstico e, consequentemente enfatizando a importância do acompanhamento a longo prazo dos pacientes com a doença.

Leishmanial antigens in the diagnosis of active lesions and ancient scars of American tegumentary leishmaniasis patients

Cutaneous biopsies (n = 94) obtained from 88 patients with American tegumentary leishmaniasis were studied by conventional and immunohistochemical techniques. Specimens were distributed as active lesions of cutaneous leishmaniasis (n = 53) (Group I), cicatricial lesions of cutaneous leishmaniasis (n = 35) (Group II) and suggestive scars of healed mucosal leishmaniasis patients (n = 6) (Group III). In addition, active cutaneous lesions of other etiology (n = 24) (Group C1) and cutaneous scars not related to leishmaniasis (n = 10) (Group C2) were also included in the protocol. Amastigotes in Group I biopsies were detected by routine histopathological exam (30.2%), imprint (28.2%), culture (43.4%), immunofluorescence (41.4%) and immunoperoxidase (58.5%) techniques; and by the five methods together (79.3%). In Group II, 5.7% of cultures were positive. Leishmanial antigen was also seen in the cytoplasm of macrophages and giant cells (cellular pattern), vessel walls (vascular pattern) and dermal nerves (neural pattern). Positive reaction was detected in 49 (92.5%), 20 (57%) and 4 (67%) biopsies of Groups I, II and III, respectively. Antigen persistency in cicatricial tissue may be related to immunoprotection or, on the contrary, to the development of late lesions. We suggest that the cellular, vascular and neural patterns could be applied in the immunodiagnosis of active and cicatricial lesions in which leishmaniasis is suspected.

Gastric cryptosporidiosis as a clue for the diagnosis of the acquired immunodeficiency syndrome

Infecções oportunistas do trato gastrointestinal constituem ameaça à população crescente de portadores de imunossupressão. O comprometimento do estômago por Cryptosporidium é incomum. Quando identificado no exame histopatológico da mucosa gástrica, é mandatória a investigação do estado imunológico do hospedeiro. São apresentados os dados clinicopatológicos e endoscópicos de uma paciente de 64 anos com gastrite erosiva associada à infecção por Cryptosporidium. O encontro deste agente oportunista no exame histopatológico da mucosa gástrica foi fundamental para esclarecer a doença de base da paciente, que era a síndrome da imunodeficiência adquirida.

Myxosporidiosis in intensively-reared Piaractus mesopotamicus: Histopathological diagnosis by means of Ziehl-Neelsen staining

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Accuracy of simple urine tests for diagnosis of urinary tract infections in low-risk pregnant women

Alterações anatômicas e fisiológicas da gravidez predispõem gestantes a infecções do trato urinário (ITU). O objetivo deste estudo foi identificar a acurácia do exame de urina simples para diagnóstico de ITU em gestantes de baixo risco. Fez-se uso do estudo de desempenho de teste diagnóstico realizado em Botucatu, SP, com 230 gestantes, entre 2006 e 2008. Os resultados mostram que a prevalência de ITU foi de 10%. A sensibilidade foi 95,6%, especificidade 63,3% e acurácia 66,5% do exame de urina simples, em relação ao diagnóstico de ITU. A análise dos valores preditivos positivo e negativo (VPP e VPN) mostrou que, na vigência de exame de urina simples normal, a chance de haver ITU foi pequena (VPN 99,2%). Frente ao resultado alterado desse exame, a probabilidade de haver ITU foi baixa (VPP 22,4%). Conclui-se que a acurácia do exame de urina simples como meio diagnóstico de ITU foi baixa, sendo indispensável a realização de urocultura para o diagnóstico.

Case for diagnosis

Descreve-se caso clínico de paciente feminina adulta, portadora de doença de Parkinson em uso de amantadina que desenvolveu extenso quadro de livedo reticular nos membros e abdome após o início do medicamento. Discutem-se a semiotécnica diagnóstica do livedo reticular e seus diferenciais. Os autores salientam que a etiologia medicamentosa deva ser considerada no diagnóstico dos livedos reticulares.

Evaluation of a novel kit (TF-test) for the diagnosis of intestinal parasitic infections

Intestinal parasitic infections are currently a source of concern for Public Health agencies in developing and developed countries. Since three ovum-and-parasite stool examinations have been demonstrated to provide sensitive results, we designed a practical and economical kit (TF-Test) that is now commercially available (Immunoassay Com. Ind. Ltda., S (a) over tildeo Paulo, Brazil). This kit allows the separate collection of three fecal specimens into a preservative solution. The specimens are then pooled, double-filtered, and concentrated by a single rapid centrifugation process. The TF-Test was evaluated in four different laboratories in a study using 1,102 outpatients and individuals living in an endemic area for enteroparasitosis. The overall sensitivity found using the TF-Test (86.2-97.8%) was significantly higher (P<0.01) than the sensitivity of conventional techniques such as the Coprotest (NIL Comercio Exterior Ltda, São Paulo, Brazil) and the combination of Lutz/Hoffman, Faust, and Rugai techniques (De Carli, Diagnostico Laboratorial das Parasitoses Humanas. Metodos e Tecnicas, 1994), which ranged from 48.3% to 75.9%. When the above combined three specimen technique was repeated with three specimens collected on different days, its sensitivity became similar (P > 0.01) to that of the TF-Test. The kappa index values of agreement for the TF-Test were consistent (P < 0.01), being higher and ranking in a better position than conventional techniques. The high sensitivity, cost/benefit ratio, and practical aspects demonstrate that the TF-Test is suitable for individual diagnosis, epidemiological inquiries, or evaluation of chemotherapy in treated communities. (C) 2004 Wiley-Liss, Inc.

Inv dup (15): Is the electroclinical phenotype helpful for this challenging clinical diagnosis?

Objective: To study the electroclinical phenotype in 5 patients with large Supernumerary marker chromosome referred as inv dup (15), in an attempt to analyze the electroclinical spectrum in order to determine if the binomial epilepsy-EEG is stereotyped enough to corroborate this challenging diagnosis.Methods: Five patients with large inv dup (15) were submitted to EEG and/or V-EEG, with a minimum duration of 2 h. Two certified neurophysiologists analyzed all EEG tracings simultaneously, blinded to clinical and molecular data. Epilepsy was characterized by detailed history and a standard questionnaire according to International League Against Epilepsy guidelines and corroborated by V-EEG findings.Results: Epilepsy started during infancy in 4 patients, in 3 with spasms. Spasms were easily controlled in one but not in others. Epilepsy evolved with generalized seizures in two patients and, generalized and focal in one. Currently, 3 patients present refractory epilepsy and two are seizure-free. In one patient, only one isolated episode suggestive of a secondary generalized tonic-clonic event occurred at the age of 12 years without recurrence. Regarding the EEG, patients had distinct features, except for two patients with very high amplitude fast activity, resembling recruiting rhythm. Despite good seizure outcome in 3 patients, EEGs remained remarkably abnormal with frequent epileptiform discharges over poorly organized background.Conclusions: Our data showed a heterogeneous electroclinical phenotype with generalized and partial epilepsy, presenting distinct degrees of severity and refractoriness.Significance: Our findings suggest that it is not possible to delineate an electroclinical phenotype in this neurogenetic syndrome. Therefore, inv dup (15) remains as a diagnostic challenge and epilepsy and EEG features are valuable only when inserted in the proper clinical context. (c) 2006 International Federation of Clinical Neurophysiology. Published by Elsevier B.V.. All rights reserved.